Development and characterization of aptamer-based enzyme-linked apta-sorbent assay for the detection of Singapore grouper iridovirus infection

Abstract

Aims: Singapore grouper iridovirus (SGIV) is a devastating aquaculture virus responsible for heavy economic losses to grouper, Epinephelus sp. aquaculture. The aim of this study was to develop a rapid and sensitive detection method for SGIV infections in infected groupers. Methods and Results: We previously generated DNA aptamers against SGIVinfected cells. In this study, we established and characterized a novel aptamer (Q3)-based enzyme-linked apta-sorbent assay (ELASA) for the detection of SGIV infection in Epinephelus coioides. The Q3-based ELASA could detect SGIV infection rapidly in vitro and in vivo, with high specificity and stability.

Molecular biology of Macrobrachium rosenbergii nodavirus infection in giant freshwater prawn

“…To date, the application of aptamers technology in infectious disease diagnosis of aquatic animal is mostly limited to viral infections. Several aptamers have been reported for viral detections (Aoki & Hirono, ; Li et al., ; Li, Zhou, et al., , Zhou et al., , ) and the detection of virus‐infected cells (Li, Wei, et al., ; Li et al., , ), and aptamers that manifesting antiviral property (Hwang et al., ; Li et al., ; Li, Zhou, et al., ; Zhou et al., ).…”

Development of novel aptamer‐based enzyme‐linked apta‐sorbent assay (ELASA) for rapid detection of mariculture pathogen Vibrio alginolyticus

“…Unfortunately, ELISA is based on polyclonal or monoclonal antibodies, which need strict low‐temperature preservation conditions required for the antibodies. Furthermore, it is rather complicated work to obtain specific antibodies up to now, let alone no way to generate antibodies against non‐immunogenic or toxic molecules (Li et al, ). Then, we chose aptamers as alternatives to antibodies in ELISA in this study.…”

“…The excellent characteristics of easy synthesis, low immunogenicity, innocuity and high specificity and stability make aptamers ideal alternatives to antibodies (Toh et al, ). Aptamer‐based enzyme‐linked apta‐sorbent assay (ELASA) has some excellent advantages than conventional ELISA: for example, aptamers used in ELASA could be obtained quickly by SELEX through in vitro chemical process, while antibodies have to selected in the biological system, which is much more time‐consuming and costly; the specificity of aptamers is reversible, and even aptamers are denatured at high temperature, which is impossible for antibodies in ELISA; and the storage conditions of ELASA are less stricter than ELISA, as the structural stability of aptamers is higher than that of antibodies (Li et al, ; Zhou et al, ). Zhou et al () have developed aptamer (A10)‐based ELASA for RGNNV diagnosis, which could accurately detect RGNNV infection at number of RGNNV‐infected cells as low as 4 × 10 3 with incubation time as short as 10 min, and A10‐ELASA results were consistent with those of reverse transcription qPCR.…”

“…The VA2‐ELASA was performed as per the protocol described previously with some modifications (Li et al, ; Toh, Citartan, Gopinath, & Tang, ). Aptamer VA2 was labelled with biotin.…”

“…There have been some techniques for identifying V. alginolyticus infection in recent years, including polymerase chain reaction (PCR) assay (Liu, Cheng, Hsu, & Chen, ), real‐time PCR (Zhou et al, ), immunofluorescence antibody test (Huang et al, ) and loop‐mediated isothermal amplification (LAMP) (Plaon, Longyant, Sithigorngul, & Chaivisuthangkura, ). However, these techniques applied to pathogen detection by researches in the laboratory but not definite diagnosis by farmers in mariculture farms because of some disadvantages, such as complicated operation, expensive reagents, time‐consuming and high required environments (Duan et al, ; Li et al, ). Therefore, novel detection techniques for the rapid diagnosis of V. alginolyticus infection in mariculture farms are urgently required.…”